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Genetic analyses of the functions of [NiFe]-hydrogenase maturation endopeptidases in the hyperthermophilic archaeon Thermococcus kodakarensis

  • Special Feature: Original Paper
  • 11th International Congress on Extremophiles
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Abstract

The maturation of [NiFe]-hydrogenases requires a number of accessory proteins, which include hydrogenase-specific endopeptidases. The endopeptidases carry out the final cleavage reaction of the C-terminal regions of [NiFe]-hydrogenase large subunit precursors. The hyperthermophilic archaeon Thermococcus kodakarensis harbors two [NiFe]-hydrogenases, a cytoplasmic Hyh and a membrane-bound Mbh, along with two putative hydrogenase-specific endopeptidase genes. In this study, we carried out a genetic examination on the two endopeptidase genes, TK2004 and TK2066. Disruption of TK2004 resulted in a strain that could not grow under conditions requiring hydrogen evolution. The Mbh large subunit precursor (pre-MbhL) in this strain was not processed at all whereas Hyh cleavage was not affected. On the other hand, disruption of TK2066 did not affect the growth of T. kodakarensis under the conditions examined. Cleavage of the Hyh large subunit precursor (pre-HyhL) was impaired, but could be observed to some extent. In a strain lacking both TK2004 and TK2066, cleavage of pre-HyhL could not be observed. Our results indicate that pre-MbhL cleavage is carried out solely by the endopeptidase encoded by TK2004. Pre-HyhL cleavage is mainly carried out by TK2066, but TK2004 can also play a minor role in this cleavage.

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Acknowledgments

This work was partially funded by the Core Research for Evolutional Science and Technology program of the Japan Science and Technology Agency to H.A. and T.I. within the research area ‘Creation of Basic Technology for Improved Bioenergy Production through Functional Analysis and Regulation of Algae and Other Aquatic Microorganisms’. The work was also partially funded to T.K. by JSPS KAKENHI (Grant Number 26292038).

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Authors and Affiliations

  1. Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Katsura, Nishikyo-ku, Kyoto, 615-8510, Japan

    Tamotsu Kanai, Ayako Yasukochi, Jan-Robert Simons, Joseph Walker Scott & Haruyuki Atomi

  2. Department of Biotechnology, College of Life Sciences, Ritsumeikan University, Kusatsu, 525-8577, Japan

    Wakao Fukuda & Tadayuki Imanaka

  3. JST, CREST, 7, Gobancho, Chiyoda-ku, Tokyo, 102-0076, Japan

    Jan-Robert Simons, Tadayuki Imanaka & Haruyuki Atomi

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  1. Tamotsu Kanai
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  2. Ayako Yasukochi
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  3. Jan-Robert Simons
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  4. Joseph Walker Scott
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  5. Wakao Fukuda
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  6. Tadayuki Imanaka
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  7. Haruyuki Atomi
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Correspondence to Haruyuki Atomi.

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Communicated by A. Driessen.

This article is part of a special feature based on the 11th International Congress on Extremophiles held in Kyoto, Japan, September 12–16, 2016.

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Kanai, T., Yasukochi, A., Simons, JR. et al. Genetic analyses of the functions of [NiFe]-hydrogenase maturation endopeptidases in the hyperthermophilic archaeon Thermococcus kodakarensis . Extremophiles 21, 27–39 (2017). https://doi.org/10.1007/s00792-016-0875-1

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